U.S. patent application number 10/857756 was filed with the patent office on 2004-12-16 for cinnamaldehyde derivatives inhibiting growth of tumor cell and regulating cell cycle, preparations and pharmaceutical compositions thereof.
Invention is credited to Choi, Sung-Gyu, Han, Dong Cho, Jeon, Sun Bok, Kim, Jong Han, Kwon, Byoung-Mog, Lee, Mi-Young, Lee, Sangku, Son, Kwang-Hee.
Application Number | 20040254196 10/857756 |
Document ID | / |
Family ID | 33509645 |
Filed Date | 2004-12-16 |
United States Patent
Application |
20040254196 |
Kind Code |
A1 |
Kwon, Byoung-Mog ; et
al. |
December 16, 2004 |
Cinnamaldehyde derivatives inhibiting growth of tumor cell and
regulating cell cycle, preparations and pharmaceutical compositions
thereof
Abstract
The present invention relates to cinnamaldehyde derivatives
inhibiting growth of tumor cell and regulating cell cycle, the
method for preparation and the pharmaceutical composition thereof.
The cinnamaldehyde derivatives of the present invention can be
effectively used as a cell cycle regulator or a cancer cell growth
inhibitor, since it has an ability to regulate cell cycle by
holding the cells in G2/M stage of the cell division and has
activity to inhibit cancer cell growth.
Inventors: |
Kwon, Byoung-Mog;
(Yusung-ku, KR) ; Son, Kwang-Hee; (Yusung-ku,
KR) ; Han, Dong Cho; (Yusung-ku, KR) ; Lee,
Sangku; (Yusung-ku, KR) ; Kim, Jong Han;
(Kyungsangnam-do, KR) ; Choi, Sung-Gyu; (Joong-ku,
KR) ; Lee, Mi-Young; (Kangwon-do, KR) ; Jeon,
Sun Bok; (Choongchungbook-do, KR) |
Correspondence
Address: |
MUSERLIAN AND LUCAS AND MERCANTI, LLP
475 PARK AVENUE SOUTH
NEW YORK
NY
10016
US
|
Family ID: |
33509645 |
Appl. No.: |
10/857756 |
Filed: |
May 28, 2004 |
Current U.S.
Class: |
514/255.03 ;
514/701; 544/392; 568/442 |
Current CPC
Class: |
C07C 47/232 20130101;
C07C 69/017 20130101; C07C 47/277 20130101; C07C 67/293 20130101;
C07C 47/24 20130101; C07C 45/72 20130101; C07D 295/112 20130101;
C07C 69/24 20130101; C07C 47/27 20130101; C07C 69/78 20130101; C07C
69/76 20130101; C07C 205/35 20130101; C07C 67/293 20130101; C07C
69/24 20130101; C07C 67/293 20130101; C07C 69/78 20130101; C07C
67/293 20130101; C07C 69/76 20130101; C07C 67/293 20130101; C07C
69/017 20130101; C07C 67/293 20130101; C07C 69/21 20130101 |
Class at
Publication: |
514/255.03 ;
514/701; 544/392; 568/442 |
International
Class: |
A61K 031/495; C07D
241/04; A61K 031/11; C07C 047/565; C07C 047/575 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 10, 2003 |
KR |
10-2003-0037198 |
Claims
What is claimed is:
1. Cinnamaldehyde derivatives represented by formula 1. 11(wherein,
R.sub.1 and R.sub.1' are same, R.sub.2 and R.sub.2' are same, and
R.sub.3 and R.sub.3'are same. R.sub.1, R.sub.2 and R.sub.3 are
independent each other, hydrogen, hydroxy, halogen,
C.sub.1.about.C.sub.4 alkyl, C.sub.1.about.C.sub.4 alkoxy,
12OCOR.sub.5, N-methylpiperazine; R.sub.4 is hydrogen, nitro,
C.sub.1.about.C.sub.4 alkoxy; R.sub.5 is C.sub.1.about.C.sub.4
alkyl, phenyl, phenyl substituted by halogen.)
2. Cinnamaldehyde derivatives as set forth in claim 1, the
cinnamaldehyde derivative is selected from the group consisting of
1) 2,3-bis-benzylidenesuccinaldehyde; 2)
2,3-bis-(2-fluorobenzylidene)succin- aldehyde; 3)
2,3-bis-(2-chlorobenzylidene)succinaldehyde; 4)
2,3-bis-(2-bromobenzylidene)succinaldehyde; 5)
2,3-bis-(2-methoxybenzylid- ene)succinaldehyde; 6)
2,3-bis-(3-chlorobenzylidene)succinaldehyde; 7)
2,3-bis-(4-hydroxy-3-methoxybenzylidene)succinaldehyde; 8)
2,3-bis-(3,4-dimethoxybenzylidene)succinaldehyde; 9)
2,3-bis-(4-chlorobenzylidene)succinaldehyde;
10)2,3-bis-(4-hydroxy-benzyl- idene)succinaldehyde;
11)2,3-bis-(4-methyl-benzylidene)succinaldehyde;
12)2,3-bis-(4-methoxy-benzylidene)succinaldehyde;
13)2,3-bis-(2-propyloxy- -benzylidene)succinaldehyde;
14)2,3-bis-(2-allyloxy-benzylidene)succinalde- hyde;
15)2,3-bis-(2-isopropyloxybenzylidene)succinaldehyde;
16)2,3-bis-(2-benzyloxy-benzylidene)succinaldehyde;
17)2,3-bis-(2-(4-chlorobenzyloxy)-benzylidene)succinaldehyde;
18)2,3-bis-(2-(4-bromobenzyloxy)-benzylidene)succinaldehyde;
19)2,3-bis-(2-(4-nitrobenzyloxy)-benzylidene)succinaldehyde;
20)2,3-bis-(3-propyloxy-benzylidene)succinaldehyde;
21)2,3-bis-(3-isopropyloxy-benzylidene)succinaldehyde;
22)2,3-bis-(3-benzyloxy-benzylidene)succinaldehyde;
23)2,3-bis-(4-propyloxy-3-methoxybenzylidene)succinaldehyde;
24)2,3-bis-(4-isopropyloxy-3-methoxybenzylidene)succinaldehyde;
25)2,3-bis-(4-acetyloxy-3-methoxybenzylidene)succinaldehyde;
26)2,3-bis-(4-valeryloxy-3-methoxybenzylidene)succinaldehyde;
27)2,3-bis-(4-benzoyloxy-3-methoxybenzylidene)succinaldehyde;
28)2,3-bis-(4-propyloxy-benzylidene)succinaldehyde;
29)2,3-bis-(4-isopropyloxybenzylidene)succinaldehyde;
30)2,3-bis-(4-valeryloxy-benzylidene)succinaldehyde;
31)2,3-bis-(4-benzyloxy-benzylidene)succinaldehyde;
32)2,3-bis-(4-benzoyloxy-benzylidene)succinaldehyde;
33)2,3-bis-(4-(2-fluorobenzoyloxy)-benzylidene)succinaldehyde;
34)2,3-bis-(4-(4-bromobenozyloxy)-benzylidene)succinaldehyde;
35)2,3-bis-(2-(N-methylpiperazine)benzylidene)succinaldehyde;
36)2,3-bis-(3-(4-chlorobenzyloxy)-benzylidene)succinaldehyde;
37)2,3-bis-(3-(4-methoxybenzyloxy)-benzylidene)succinaldehyde;
38)2,3-bis-(4-(4-chlorobenzyloxy)-benzylidene)succinaldehyde; and
39)2,3-bis-(4-(4-methoxybenzyloxy)-benzylidene)succinaldehyde.
3. The method for preparation of the cinnamaldehyde
derivatives(II), the cinnamaldehyde derivatives are prepared by
linear dimerization of benzaldehyde substituted by R.sub.1, R.sub.2
and R.sub.3(represented by formula I) in the presence of
2,5-dimethoxytetrahydrofuran, potassium acetate, acetic acid and
water. 13(wherein, R.sub.1, R.sub.1', R.sub.2, R.sub.2', R.sub.3
and R.sub.3' are described in the formula 1)
4. The pharmaceutical composition for inhibiting growth of tumor
cell comprising the cinnamaldehyde derivative of claim 1 as
effective ingredient.
5. The pharmaceutical composition for regulating cell cycle
comprising the cinnamaldehyde derivative of claim 1 as effective
ingredient.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates to cinnamaldehyde derivatives
represented by formula 1, process for preparation and
pharmaceutical compositions thereof. Particularly, the present
invention relates to cinnamaldehyde derivatives inhibiting growth
of tumor cell and regulating cell cycle, the method for preparation
thereof and the pharmaceutical composition as tumor cell growth
inhibitor or cell cycle regulator. 1
[0003] (wherein, R.sub.1, R.sub.1', R.sub.2, R.sub.2', R.sub.3 and
R.sub.3' are described in the below.)
[0004] 2. Description of the Prior Art
[0005] Proliferation, differentiation and apoptosis are the major
phenomena to keep a life. In order for cell to function normally,
cellular proliferation, differentiation, and apoptosis are
regulated by an elaborate intracellular and intercellular signal
transduction system. That is, once cells are given a signal from
outside, the signal is transferred to cellular clock by signaling
proteins (PLC, PKC, Shc, Grb2, Raf, MAPK, MEK, etc.) and signaling
messengers (GTP, cAMP, etc.). Any abnormality in the process causes
diseases such as cancer.
[0006] Cell cycle is composed of following stages; Gap(G1), DNA
synthesis(S), Gap2(G2) and Meiosis(M). In addition to the stages,
when cells have been under the condition of having lower
concentration of a growth factor for a long while, the cells get
into the resting stage(Go), in which cell growth is stopped.
[0007] In cell cycle, a very complicated network, so called `check
point`, makes the cell clock move properly in the order of
G1-S-G2-M. The obstruction of the regulating mechanism of the check
point results in uncontrolled cell growth.
[0008] If signal transduction from outside of the nucleus is smooth
and nutritional condition is good, cells become larger in stage G1
and then entered cell cycle. Cell cycle goes into action in G1
check point which is named as start point in yeast cells, and
restriction point in mammalian cells. After passing through the
stage, if there is no specific obstruction, cells go through the 4
stages automatically, leading to the replication of genomes and
differentiation. The procedure, especially in mammalian cells, is
precisely explained hereinafter. The stage G1 having the check
point is a preparatory stage for making new cells. At this time,
enough growth factors and nutrition should be given to cells.
Otherwise, cell cycle is stopped and cells go into the stage Go
with no more growth. However, cell cycle progresses to the stage S
under the supply of various growth factors and nutrition. At this
time, replication of genome is carried out, two copies of
chromosomes are produced, and various factors in cytoplasm are
duplicated as well in order for a cell to be differentiated into
two individual cells. After passing through the stage S, cells go
into the stage G2, which can be said as the second check point.
During the stage G2, DNA replication is regulated and completed,
and entry to meiosis (M stage) is prepared. Lots of factors
essential for the construction of a cell are generated in this
stage. After the generation of all the required factors for the
cell division, cells progress to the stage M, the stage in which
actual cell proliferation occurs. The stage M has the shortest
period among stages. In this stage, the duplicated genome is
separated and each part moves to both poles, resulting in two
daughter cells. All the stages are required in order for a cell to
divide into two cells, so that they are very important for
continuing the life of a cell. Thus, the studies on cell cycle and
the development of a regulator of the cell cycle are prerequisite
for the studies on mechanisms of cell growth and for the
development of a preventive or a treatment agent for cancer caused
by the abnormality of cell cycle (Nature Review Cancer, 2001, 1,
222-231).
[0009] As mentioned above, mammalian cell growth can be regulated
by controlling the first check point in G1 stage or the second
check point in G2/M stage. The abnormal progress of the first or
the second check point is involved in cellular ageing or the
development of cancer. And cycline D (D1, D2, D3, etc.) plays an
important role in those check points. Cycline D regulates the
enzyme activity by being combined with cycline dependant kinases
(CDK; CDK2, CDK4, CDK6), and is also deeply involved in the
regulation of whole cell cycle by CDC25 which functions to remove
phosphate group of a phosphorylated protein. Based on that
founding, it is no wonder that various cell cycle regulators have
been good candidates for the development of a treatment agent for
intractable diseases such as cancers. (Current Opinion in Chemical
Biology, 2002, 6, 472-278).
[0010] Thus, the present inventors have disclosed that the novel
cinnamaldehyde derivatives could block the G2/M selectively and
have completed the invention by confirming that the cinnamaldehyde
derivatives could be effectively used as a cell cycle regulator and
as an inhibitor for abnormal cell growth of cancer cells.
OBJECT OF THE INVENTION
[0011] It is an object of the present invention to provide
cinnamaldehyde derivatives used as tumor cell growth inhibitor or
cell cycle regulator by selectively holding cells in G2/M of cell
cycle.
[0012] It is another object of the present invention to provide the
method for preparation the cinnamaldehyde derivatives.
[0013] It is a further object of the present invention to provide
the pharmaceutical compositions comprising cinnamaldehyde
derivatives as effective ingredient.
SUMMARY OF THE INVENTION
[0014] The present invention provides cinnamaldehyde derivatives
represented by formula 1. 2
[0015] (wherein, R.sub.1 and R.sub.1' are same, R.sub.2 and
R.sub.2' are same, and R.sub.3 and R.sub.3'are same.
[0016] R.sub.1, R.sub.2 and R.sub.3 are independent each other,
hydrogen, hydroxy, halogen, C.sub.1.about.C.sub.4 alkyl,
C.sub.1.about.C.sub.4 alkoxy, 3
[0017] OCOR.sub.5, N-methylpiperazine;
[0018] R.sub.4 is hydrogen, nitro, C.sub.1.about.C.sub.4
alkoxy;
[0019] R.sub.5 is C.sub.1.about.C.sub.4 alkyl, phenyl, phenyl
substituted by halogen.)
[0020] The present invention preferably provides compounds
represented by the followings.
[0021] 1) 2,3-bis-benzylidenesuccinaldehyde;
[0022] 2) 2,3-bis-(2-fluorobenzylidene)succinaldehyde;
[0023] 3) 2,3-bis-(2-chlorobenzylidene)succinaldehyde;
[0024] 4) 2,3-bis-(2-bromobenzylidene)succinaldehyde;
[0025] 5) 2,3-bis-(2-methoxybenzylidene)succinaldehyde;
[0026] 6) 2,3-bis-(3-chlorobenzylidene)succinaldehyde;
[0027] 7)
2,3-bis-(4-hydroxy-3-methoxybenzylidene)succinaldehyde;
[0028] 8) 2,3-bis-(3,4-dimethoxybenzylidene)succinaldehyde;
[0029] 9) 2,3-bis-(4-chlorobenzylidene)succinaldehyde;
[0030] 10)2,3-bis-(4-hydroxy-benzylidene)succinaldehyde;
[0031] 11)2,3-bis-(4-methyl-benzylidene)succinaldehyde;
[0032] 12)2,3-bis-(4-methoxy-benzylidene)succinaldehyde;
[0033] 13)2,3-bis-(2-propyloxy-benzylidene)succinaldehyde;
[0034] 14)2,3-bis-(2-allyloxy-benzylidene)succinaldehyde;
[0035] 15)2,3-bis-(2-isopropyloxybenzylidene)succinaldehyde;
[0036] 16)2,3-bis-(2-benzyloxy-benzylidene)succinaldehyde;
[0037]
17)2,3-bis-(2-(4-chlorobenzyloxy)-benzylidene)succinaldehyde;
[0038]
18)2,3-bis-(2-(4-bromobenzyloxy)-benzylidene)succinaldehyde;
[0039]
19)2,3-bis-(2-(4-nitrobenzyloxy)-benzylidene)succinaldehyde;
[0040] 20)2,3-bis-(3-propyloxy-benzylidene)succinaldehyde;
[0041] 21-)2,3-bis-(3-isopropyloxybenzylidene)succinaldehyde;
[0042] 22)2,3-bis-(3-benzyloxy-benzylidene)succinaldehyde;
[0043]
23)2,3-bis-(4-propyloxy-3-methoxybenzylidene)succinaldehyde;
[0044]
24)2,3-bis-(4-isopropyloxy-3-methoxybenzylidene)succinaldehyde;
[0045]
25)2,3-bis-(4-acetyloxy-3-methoxybenzylidene)succinaldehyde;
[0046]
26)2,3-bis-(4-valeryloxy-3-methoxybenzylidene)succinaldehyde;
[0047]
27)2,3-bis-(4-benzoyloxy-3-methoxybenzylidene)succinaldehyde;
[0048] 28)2,3-bis-(4-propyloxy-benzylidene)succinaldehyde;
[0049] 29)2,3-bis-(4-isopropyloxybenzylidene)succinaldehyde;
[0050] 30)2,3-bis-(4-valeryloxy-benzylidene)succinaldehyde;
[0051] 31)2,3-bis-(4-benzyloxy-benzylidene)succinaldehyde;
[0052] 32)2,3-bis-(4-benzoyloxy-benzylidene)succinaldehyde;
[0053]
33)2,3-bis-(4-(2-fluorobenzoyloxy)-benzylidene)succinaldehyde;
[0054]
34)2,3-bis-(4-(4-bromobenozyloxy)-benzylidene)succinaldehyde;
[0055]
35)2,3-bis-(2-(N-methylpiperazine)benzylidene)succinaldehyde;
[0056]
36)2,3-bis-(3-(4-chlorobenzyloxy)-benzylidene)succinaldehyde;
[0057]
37)2,3-bis-(3-(4-methoxybenzyloxy)-benzylidene)succinaldehyde;
[0058]
38)2,3-bis-(4-(4-chlorobenzyloxy)-benzylidene)succinaldehyde;
and
[0059]
39)2,3-bis-(4-(4-methoxybenzyloxy)-benzylidene)succinaldehyde.
[0060] The compound of the present invention shown in the above
formula 1 can be effectively used as a cancer cell growth inhibitor
or a cell cycle regulator, since it regulates cell cycle by holding
a cell in the middle of division in stage G2/M of cell cycle and
inhibits the cancer cell growth thereby.
[0061] The present invention provides the method for preparation of
the cinnamaldehyde derivatives represented by formula 1.
[0062] As shown in scheme 1, the cinnamaldehyde derivatives of the
present invention are prepared by linear dimerization of
benzaldehyde substituted by R.sub.1, R.sub.2 and
R.sub.3(represented by formula I). Particularly, as shown in scheme
1, the cinnamaldehyde derivatives(II) of the present invention are
prepared by linear dimerization of benzaldehyde substituted by
R.sub.1, R.sub.2 and R.sub.3(represented by formula I) in the
presence of 2,5-dimethoxytetrahydrofuran, potassium acetate, acetic
acid and water. 4
[0063] (wherein, R.sub.1, R.sub.1', R.sub.2, R.sub.2', R.sub.3 and
R.sub.3' are described in the formula 1)
[0064] The benzaldehydes substituted with R.sub.1, R.sub.2 and
R.sub.3(represented by formula I) are used as commercially
available formulation. Also, the benzaldehydes substituted with
R.sub.1, R.sub.2 and R.sub.3 (represented by formula I) are
prepared by reacting alkylating agent to benzaldehyde substituted
with activating substitutent such as halogen or hydroxy group in
the presence of base, but not is limited to them. As shown in
example 13, for example, 2-propyloxybenzaldehyde(I) was prepared by
reacting 1-iodopropane(alkylating agent) to 2-hydroxybenzaldehyde
substituted with hydroxyl group in the presence of potassium
carbonate(base).
[0065] Also, the cinnamaldehyde derivatives(II) of the present
invention are prepared by dimerizing the benzaldehyde (by linear
dimerization) and transforming R.sub.1, R.sub.1', R.sub.2,
R.sub.2', R.sub.3 and R.sub.3'. Then, R.sub.1, R.sub.1', R.sub.2,
R.sub.2', R.sub.3 and R.sub.3' are transformed by reacting the
dimerized product to alkylating agent in the presence of base, but
not is limited to them. As shown in example 23, for example,
2,3-bis-(4-hydroxy-3-methoxybenzylidene) was prepared by
linear-dimerizing vaniline in the presence of
2,5-dimethoxytetrahydrofura- n, potassium acetate, acetic acid and
water, thereafter the dimerized product reacted
1-iodopropan(alkylating agent) in the presence of potassium
acetate(base) to transform hydroxy group to propyloxy group.
[0066] Also, the present invention provides the pharmaceutical
compositions as tumor cell growth inhibitor or cell cycle
regulator, comprising cinnamaldehyde derivatives as effective
ingredient.
[0067] The pharmaceutical compositions comprising cinnamaldehyde
derivatives as effective ingredient can be administered through
various administration routes such as oral or parenteral
formulations in practically clinical testing. Also, the
pharmaceutical compositions can be used as medicine.
[0068] The cinnamaldehyde derivatives of the present invention can
be administered to a human body through various formulations.
[0069] The pharmaceutical compositions of the present invention
contain further the pharmaceutically acceptable carrier. More
particularly, the pharmaceutically acceptable carrier is any of the
standard pharmaceutical carriers used in the known formulations,
such as sterile solution, tablet, coating tablet and capsule.
Conservatively, the carrier is selected from the group of excipient
such as starch, milk, glucose, specific clay, gelatin, stearic
acid, talc, vegetable oil or fat, gum, glycol, the other known
excipients, flavoring agents, pigment additives and other
components.
[0070] Pharmaceutical compositions as tumor cell growth inhibitor
or cell cycle regulator, containing cinnamaldehyde derivatives of
the formula 1 according to the present invention is administered,
but not is limited, through conservative routes such as oral,
intravenous injection, intramuscular injection, transdermal
administration. For example, the cinnamaldehyde derivatives
according to the present invention can be administered to a human
body through various oral or parenteral formulations in practically
clinical testing. Formulations are prepared by using available
additives such as packing agents, bulking agents, binding agents,
disintegrants and surfactants, or excipients. Solid formulations
for oral administration are provided into various forms including
tablets, pills, powders, granules and capsules. Solid formulations
are prepared by mixing one or more compounds selected from the
group consisting of cinnamaldehyde derivatives of the formula 1,
and at least one excipient which is selected from the group
consisting of starch, calcium carbonate, sucrose or lactose, and
gelatin. Also, lubricants such as magnesium stereate talc can be
used together with simple diluting agent. Liquid formulations for
oral administration are provided into suspension, solution,
emulsion and syrup. Various excipients, for example, moistening
agent, sweeting agent, aromatic agent and preservative can be
included in liquid formulations, together with simple diluting
agent, commercially available, such as water, or liquid
paraffin.
[0071] Also, the pharmaceutical composition of the present
invention can be administered to a human body parenterally.
Parenteral administration is carried out by hypodermic injection,
intravenous injection or intramuscular injection. The formulation
for parenteral administration is prepared by mixing the
cinnamaldehyde derivatives with stabilizing agents or buffering
agents in water, formulating solution or suspension and preparing
an unit dosage form, such as ample or vial.
[0072] In accordance with the present invention, cinnamaldehyde
derivatives of formula 1 are contained in the pharmaceutical
compositions in broad ranges. The dose of the medically effective
component in accordance with the present invention is selected
according to absorptivity of the active component in vivo,
activity, excretion rate, age, sex and state of the patients,
seriousness of disease under treatment. Generally, the effective
component can be administered to a body one time or many times a
day, preferably 10.about.20 mg, more preferably 5.about.10 mg.
Accurate dose, administration route and frequency of the above
preparation are dependant on property of the preparation, weight or
state of the administrative human, and property of specific
derivatives.
DETAILED DESCRIPTION OF THE INVENTION
[0073] The present invention will be explained in more detail with
reference to the following examples. However, the following
examples are provided only to illustrate the present invention, and
the present invention is not limited to them.
EXAMPLE 1
2,3-bis-benzylidenesuccinaldehyde
[0074] 5
[0075] 15.9 ml of benzaldehyde (150 mmol) was added to the 250 ml
round-bottomed flask. Thereafter 10 ml of
2,5-dimethoxytetrahydrofuran(75 mmol), 10 g of potassium acetate
(100 mmol), 5 ml of acetic acid(80 mmol) and 5 ml of water were
added to the flask. The reaction mixture was refluxed at
110.degree. C. for 12 hours. After the reaction was terminated, the
reaction mixture was cooled to the room temperature. Thereafter,
water was added to the reaction mixture. The reaction mixture was
extracted with 100 ml of chloroform for three times. The chloroform
layers were collected, washed with water for three times and dried
with anhydrous magnesium sulfate. The chloroform was removed under
reduced pressure. The resulting residue was purified with column
chromatography, to give the title compound as a yellow crystal
(yield: 10%)
[0076] mp:109.about.110.degree. C. R.sub.f:0.5 (Hexane:Ethyl
acetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm): .delta.1.08(t, 6H,
--CH.sub.3, J=7.5 Hz), 1.86(m, 4H, --CH.sub.2--, J=7.5 Hz), 3.96(m,
4H, CH.sub.2--, J=6.6 Hz), 6.73-7.44(m, 8H), 8.12(s, 2H,
CH.dbd.C--), 9.66(s, 2H, CH.dbd.O)
EXAMPLE 2.about.12
[0077] In the Example 1, benzaldehyde substituted with R.sub.1,
R.sub.2 and R.sub.3 shown in table 1 was used in place of
benzaldehyde, and the same procedure as the example 1 was
accomplished.
EXAMPLE 2
2,3-bis-(2-fluorobenzylidene)succinaldehyde
[0078] mp:141.about.142.degree. C. Rf=0.35
(Hexane:Ethylacetate=6:4). .sup.1 H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.7.00.about.7.08(d, 4H.sub.arom), 7.29.about.7.41(m,
4H.sub.arom), 7.88(s, 2H, CH.dbd.C--), 9.69(s, 2H, CH.dbd.O).
EXAMPLE 3
2,3-bis-(2-chlorobenzylidene)succinaldehyde
[0079] mp:177.about.178.degree. C. Rf=0.31
(Hexane:Ethylacetate=6:4) .sup.1H-NMR (CDCl.sub.3/TMS) (ppm):
.delta.7.13.about.7.28 (m, .sup.8H.sub.arom), 7.83 (s, 2H,
CH.dbd.C--), 9.73(s, 2H, CH.dbd.O).
EXAMPLE 4
2,3-bis-(2-bromobenzylidene)succinaldehyde
[0080] mp:194.about.195.degree. C. Rf=0.29
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.7.12.about.7.46(m, 8H.sub.arom), 7.73(s, 2H, CH.dbd.C--),
9.74(s, 2H, CH.dbd.O).
EXAMPLE 5
2,3-bis-(2-methoxybenzylidene)succinaldehyde
[0081] mp:163.about.164.degree. C. Rf=0.25
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.3.85 (s, 6H, --OCH.sub.3), 6.76.about.7.41(m, 8H.sub.arom),
8.07(s, 2H, CH.dbd.C--), 9.65(s, 2H, CH.dbd.O).
EXAMPLE 6
2,3-bis-(3-chlorobenzylidene)succinaldehyde
[0082] mp:107.about.108.degree. C. Rf=0.31
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.7.24.about.7.42 (m, 8H.sub.arom), 7.64 (s, 2H, CH.dbd.C--),
9.67(s, 2H, CH.dbd.O).
EXAMPLE 7
2,3-bis-(4-hydroxy-3-methoxy-benzylidene)succinaldehyde
[0083] mp:177.about.178.degree. C. Rf=0.15
(Hexane:Ethylacetate=4:6). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.3.75(s, 6H, --OCH.sub.3), 5.92(s, 2H, --OH),
6.85.about.7.16(m, 6H.sub.arom), 7.63(s, 2H, CH.dbd.C--), 9.63(s,
2H, CH.dbd.O).
EXAMPLE 8
2,3-bis-(3,4-dimethoxybenzylidene)succinaldehyde
[0084] mp:149.about.150.degree. C. Rf=0.2
(Hexane:Ethylacetate=4:6). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
6.73(s, 6H, --OCH.sub.3), 3.88(s, 6H, --OCH.sub.3),
6.81.about.7.21(m, 6H.sub.arom), 7.65(s, 2H, CH.dbd.C--), 9.65(s,
2H, CH.dbd.O).
EXAMPLE 9
2,3-bis-(4-chlorobenzylidene)succinaldehyde
[0085] mp:172.about.173.degree. C. Rf=0.31
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.7.27.about.7.44(m, 8H.sub.arom), 7.66(s, 2H, CH.dbd.C--),
9.65(s, 2H, CH.dbd.O).
EXAMPLE 10
2,3-bis-(4-hydroxy-benzylidene)succinaldehyde
[0086] mp:244.about.246.degree. C. Rf=0.25
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.6.73(dd, 4H), 7.48(dd, 4H), 7.73(s, 2H, CH.dbd.C--), 9.57(s,
2H, CH.dbd.O).
EXAMPLE 11
2,3-bis-(4-methyl-benzylidene)succinaldehyde
[0087] mp:177.about.178.degree. C. Rf=0.5
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.2.32 (s, 6H, --CH.sub.3), 7.11(dd, 4H), 7.43(dd, 4H),
7.67(s, 2H, CH.dbd.C--), 9.64(s, 2H, CH.dbd.O).
EXAMPLE 12
2,3-bis-(4-methoxy-benzylidene)succinaldehyde
[0088] mp:207.about.207.degree. C. Rf=0.23
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.3.79(s, 6H, --OCH.sub.3), 6.83(dd, 4H), 7.51(dd, 4H),
7.64(s, 2H, CH.dbd.C--), 9.63(s, 2H, CH.dbd.O).
EXAMPLE 13
2,3-bis(2-propyloxy-benzylidene)succinaldehyde
[0089] 6
[0090] (Step 1): Preparation of 2-propyloxy benzaldehyde
[0091] 6.1 g of 2-hydroxybenzaldehyde (50 mmol) was added to the
250 ml round-bottomed flask, 50 ml of acetonitrile was added
therein. Thereafter, 8.28 g of potassium carbonate (60 mmol) and
9.35 g of 1-iodopropan (55 mmol) were added therein. The reaction
mixture was refluxed at 85.degree. C. for 12 hours.
[0092] After the reaction was terminated, the reaction mixture was
cooled to the room temperature. Thereafter, water was added to the
reaction mixture. The reaction mixture was extracted with 100 ml of
chloroform for three times. The chloroform layers were collected,
washed with water for three times and dried with anhydrous
magnesium sulfate. The chloroform was removed under reduced
pressure. The resulting residue was purified with column
chromatography. Substitution of propyl group was confirmed by
hydrogen NMR.
[0093] (Step 2):Preparation of
2,3-bis(2-propyloxy-benzylidene)succinaldeh- yde
[0094] 5 g of 2-propyloxy benzaldehyde (30 mmol) prepared in the
above step 1 was added to the 250 ml round-bottomed flask.
Thereafter, 2 ml of 2,5-dimethoxytetrahydrofuran(15 mmol), 2 g of
potassium acetate (20 mmol), 1 ml of acetic acid(16 mmol) and 1 ml
of water were added to the flask. The reaction mixture was refluxed
at 110.degree. C. for 12 hours. After the reaction was terminated,
the reaction mixture was cooled to the room temperature.
Thereafter, water was added to the reaction mixture. The reaction
mixture was extracted with 100 ml of chloroform for three times.
The chloroform layers were collected, washed with water for three
times and dried with anhydrous magnesium sulfate. The chloroform
was removed under reduced pressure. The resulting residue was
purified with column chromatography, to give the title compound as
a yellow crystal (yield: 10%)
[0095] mp:109.about.110.degree. C. R.sub.f: 0.5 (Hexane:Ethyl
acetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm): .delta.1.08(t, 6H,
--CH.sub.3, J=7.5 Hz), 1.86(m, 4H, --CH.sub.2--, J=7.5 Hz), 3.96(m,
4H, CH.sub.2--, J=6.6 Hz), 6.73.about.7.44(m, 8H), 8.12(s, 2H,
CH.dbd.C--), 9.66(s, 2H, CH.dbd.O)
EXAMPLE 14.about.19
[0096] In the Example 13, alkylating agent containing R.sub.1 shown
in table 1 was used in place of 1-iodopropan, and the same
procedure as the example 13 was accomplished.
EXAMPLE 14
2,3-bis-(2-allyloxy-benzylidene)succinaldehyde
[0097] mp:117.about.118.degree. C. Rf=0.46
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.4.57(dd, 4H, CH.sub.2--CH), 5.36(q, 4H, CH.dbd.CH.sub.2),
6.07(m, 2H, C.sub.H.dbd.CH.sub.2), 6.76.about.7.42(m, 8H.sub.arom),
8.11(s, 2H, CH.dbd.C--), 9.67(s, 2H, CH.dbd.O).
EXAMPLE 15
2,3-bis-(2-isopropyloxy-benzylidene)succinaldehyde
[0098] mp:109.about.110.degree. C. Rf=0.45
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.1.38(dd, 12H, CH.sub.3--CH--CH.sub.3), 4.58(m. 2H,
CH.sub.3--CH--CH.sub.3), 6.72.about.7.44(m, 8H.sub.arom), 8.10(s,
2H, CH.dbd.C--), 9.66(s, 2H, CH.dbd.O).
EXAMPLE 16
2,3-bis-(2-benzyloxy-benzylidene)succinaldehyde
[0099] mp:120.about.121.degree. C. Rf=0.48
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.5.09(dd, 4H, CH.sub.2-Arom), 6.76.about.7.45(m,
18H.sub.arom), 8.13(s, 2H, CH.dbd.C--), 9.65(s, 2H, CH.dbd.O).
EXAMPLE 17
2,3-bis-(2-(4-chlorobenzyloxy)-benzylidene)succinaldehyde
[0100] mp:186.about.187.degree. C. Rf=0.35
(Hexane:Ethylacetate=7:3). .sup.1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.5.02(dd, 4H, CH.sub.2-Arom), 6.70.about.7.36(m,
16H.sub.arom), 8.01(s, 2H, CH.dbd.C--), 9.56(s, 2H, CH.dbd.O).
EXAMPLE 18
2,3-bis-(2-(4-bromobenzyloxy)-benzylidene)succinaldehyde
[0101] mp:210.about.211.degree. C. Rf=0.4
(Hexane:Ethylacetate=7:3). .sup.1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.5.07(dd, 4H, CH.sub.2-Arom), 6.77.about.7.54(m,
16H.sub.arom), 8.08(s, 2H, CH.dbd.C--), 9.63(s, 2H, CH.dbd.O).
EXAMPLE 19
2,3-bis-(2-(4-nitrobenzyloxy)-benzylidene)succinaldehyde
[0102] mp:116.about.117.degree. C. Rf=0.4
(Hexane:Ethylacetate=7:3). .sup.1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.0.31(s, 4H, CH.sub.2-Arom), 6.99.about.8.30(m, 16H,
H.sub.arom), 10.56(s, 2H, CH.dbd.O).
EXAMPLE 20
2,3-bis-(3-propyloxy-benzylidene)succinaldehyde
[0103] 7
[0104] (Step 1): Preparation of 3-propyloxy benzaldehyde
[0105] 6.1 g of 3-hydroxy benzaldehyde (50 mmol) was added to the
250 ml round-bottomed flask, 50 ml of acetonitrile was added
therein. Thereafter, 8.28 g of potassium carbonate (60 mmol) and
9.35 g of 1-iodopropan (55 mmol) were added therein. The reaction
mixture was refluxed at 85.degree. C. for 10 hours.
[0106] After the reaction was terminated, the reaction mixture was
cooled to the room temperature. Thereafter, water was added to the
reaction mixture. The reaction mixture was extracted with 100 ml of
chloroform for three times. The chloroform layers were collected,
washed with water for three times and dried with anhydrous
magnesium sulfate. The chloroform was removed under reduced
pressure. The resulting residue was purified with column
chromatography. Substitution of propyl group was confirmed by
hydrogen-NMR.
[0107] (Step 2):Preparation of
2,3-bis(3-propyloxy-benzylidene)succinaldeh- yde
[0108] 5 g of 3-propyloxy benzaldehyde (30 mmol) prepared in the
above step 1 was added to the 250 ml round-bottomed flask.
Thereafter, 2 ml of 2,5-dimethoxytetrahydrofuran(15 mmol), 2 g of
potassium acetate (20 mmol), 1 ml of acetic acid(16 mmol) and 1 ml
of water were added to the flask. The reaction mixture was refluxed
at 110.degree. C. for 12 hours. After the reaction was terminated,
the reaction mixture was cooled to the room temperature.
Thereafter, water was added to the reaction mixture. The reaction
mixture was extracted with 100 ml of chloroform for three times.
The chloroform layers were collected, washed with water for three
times and dried with anhydrous magnesium sulfate. The chloroform
was removed under reduced pressure. The resulting residue was
purified with column chromatography, to give the title compound as
a brown powder (1 g, yield: 10%)
[0109] mp: 93.about.94.degree. C. Rf: 0.52
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.0.99(t, 6H, --CH.sub.2--CH.sub.3, J=7.2 Hz), 1.76(m, 4H,
CH.sub.2--CH.sub.2--, J=6.9 Hz), 3.81(m, 4H, CH.sub.2--CH.sub.2,
J=l.2 Hz), 6.88.about.7.26(m, 8H arom), 7.67(s, 2H, CH.dbd.C--),
9.65(s, 2H, CH.dbd.O).
EXAMPLE 21.about.22
[0110] In the Example 20, alkylating agent containing R.sub.2 shown
in table 1 was used in place of 1-iodopropan, and the same
procedure as the example 20 was accomplished.
EXAMPLE 21
2,3-bis-(3-isopropyloxy-benzylidene)succinaldehyde
[0111] mp:116.about.117.degree. C. Rf=0.48
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.1.27(q, 12H, CH--CH.sub.3), 4.35.about.4.43(m, 2H,
CH--CH.sub.3), 6.86.about.7.26(m, 8H.sub.arom), 7.66(s, 2H,
CH.dbd.C--), 9.65(s, 2H, CH.dbd.O).
EXAMPLE 22
2,3-bis-(3-benzyloxy-benzylidene)succinaldehyde
[0112] mp:126.about.127.degree. C. Rf=0.44
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.4.97(s, 4H, CH.sub.2-ph), 6.97.about.7.38(m, 18H.sub.arom),
7.51(s, 2H, CH.dbd.C--), 9.48(s, 2H, CH.dbd.O).
EXAMPLE 23
2,3-bis-(4-propyloxy-3-methoxy benzylidene)succinaldehyde
[0113] 8
[0114] (Step 1):Preparation of
2,3-bis-(4-hydroxy-3-methoxy-benzylidene)su- ccinaldehyde
[0115] 45.65 g of vaniline (300 mmol) was added to the 250 ml
round-bottomed flask. Thereafter, 20 ml of
2,5-dimethoxytetrahydrofuran(1- 50 mmol), 20 g of potassium
acetate, 10 ml of acetic acid and 10 ml of water were added
therein. The reaction mixture was refluxed at 110.degree. C. for 12
hours.
[0116] After the reaction was terminated, the reaction mixture was
cooled to the room temperature. Thereafter, water was added to the
reaction mixture. The reaction mixture was extracted with 100 ml of
chloroform for three times. The chloroform layers were collected,
washed with water for three times and dried with anhydrous
magnesium sulfate. The chloroform was removed under reduced
pressure. The resulting residue was purified with column
chromatography. Substitution of propyl group was confirmed by
hydrogen-NMR.
[0117] (Step 2):Preparation of
2,3-bis(4-propyloxy-3-methoxybenzylidene)su- ccinaldehyde
[0118] 30 mg of
2,3-bis-(4-hydroxy-3-methoxy-benzylidene)succinaldehyde(0.- 08
mmol) prepared in the above step 1 was added to the 100 ml
round-bottomed flask. Thereafter, 30 mg of potassium carbonate(0.18
mmol) and 30 mg of 1-iodopropan(0.18 mmol) were added to the flask.
Thereafter, the mixture was stirred at 85.degree. C. for 12 hours.
After the termination of the reaction is confirmed with thin layer
chromatography, the reaction mixture was filtered to obtain the
filtrate. The residue obtained by concentrating the filtrate was
purified with silica gel column chromatography to prepare 20 mg of
the title compound as yellow powder.
[0119] mp: 95.about.96.degree. C. Rf: 0.15 (Hexane:Ethyl
acetate=4:6). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm): .delta.1.01(t, 6H,
--CH.sub.2--CH.sub.3, J=7.2 Hz), 1.80.about.1.87(m, 5H,
CH.sub.2--CH.sub.3, J=6.9 Hz), 3.70(s, 6H, --OCH.sub.3), 3.97(t,
4H, --OCH.sub.2--CH.sub.2, J=6.6 Hz), 6.78.about.7.17(m, 6H),
7.63(s, 2H, CH.dbd.C--), 9.64(s, 2H, CH.dbd.O).
EXAMPLE 24.about.27
[0120] In the Example 23, alkylating agent containing R.sub.3 shown
in table 1 was used in place of 1-iodopropan, and the same
procedure as the example 23 was accomplished.
EXAMPLE 24
2,3-bis- (4-isopropyloxy-3-methoxy-benzylidene)succinaldehyde
[0121] mp:119.about.120.degree. C. Rf=0.4
(Hexane:Ethylacetate=4:6). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.1.36(d, 12H, CH.sub.3--CH--CH.sub.3), 3.69(s, 6H,
--OCH.sub.3), 4.55.about.4.59(m, 2H,
CH.sub.3--CH--CH.sub.3),6.79.about.7.17(m, 6H.sub.arom), 7.63(s,
2H, CH.dbd.C--), 9.64(s, 2H, CH.dbd.O).
EXAMPLE 25
2,3-bis-(4-acetyloxy-3-methoxy-benzylidene)succinaldehyde
[0122] mp:149.about.151.degree. C. Rf=0.42
(Hexane:Ethylacetate=4:6). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.2.30(s, 6H, O.dbd.C--CH.sub.3), 3.72(s, 6H, --OCH.sub.3),
7.00.about.7.14(m, 6H.sub.arom), 7.68(s, 2H, CH.dbd.C--), 9.66(s,
2H, CH.dbd.O).
EXAMPLE 26
2,3-bis-(4-valeryloxy-3-methoxy-benzylidene)succinaldehyde
[0123] mp: gel Rf=0.3 (Hexane:Ethylacetate=4:6).
.sup.1H-NMR(CDCl.sub.3/TM- S) (ppm): .delta.0.96(t, 6H,
CH.sub.2--CH.sub.3), 1.38.about.1.51(m, 4H, CH.sub.2--CH.sub.3),
1.64.about.1.78(m, 4H, CH.sub.2--CH.sub.2), 2.57(t, 4H,
OCH.sub.2--CH.sub.2), 3.71(s, 6H, --OCH.sub.3), 6.99.about.7.14(m,
6H.sub.arom), 7.27(s, 2H, CH.dbd.C--), 9.65(s, 2H, CH.dbd.O).
EXAMPLE 27
2,3-bis-(4-benzoyloxy-3-methoxy-benzylidene)succinaldehyde
[0124] mp:107.about.110.degree. C. Rf=0.45
(Hexane:Ethylacetate=4:6). 1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.3.73(s, 6H, --OCH3), 7.14.about.8.82(m, 16H.sub.arom),
7.74(s, 2H, CH.dbd.C--), 9.69(s, 2H, CH.dbd.O).
EXAMPLE 28
2,3-bis-(4-propyloxybenzylidene)succinaldehyde
[0125] 9
[0126] (Step 1): Preparation of
2,3-bis-(4-hydroxybenzylidene)succinaldehy- de
[0127] 3.66 g of 4-hydroxybenzaldehyde (30 mmol) was added to the
250 ml round-bottomed flask. Thereafter, 2 ml of
2,5-dimethoxytetrahydrofuran (15 mmol), 2 g of potassium acetate, 1
ml of acetic acid and 1 ml of water were added therein. The
reaction mixture was refluxed at 110.degree. C. for 12 hours.
[0128] After the reaction was terminated, the reaction mixture was
cooled to the room temperature. Thereafter, water was added to the
reaction mixture. The reaction mixture was extracted with 100 ml of
chloroform for three times. The chloroform layers were collected,
washed with water for three times and dried with anhydrous
magnesium sulfate. The chloroform was removed under reduced
pressure. The resulting residue was purified with column
chromatography. Substitution of propyl group was confirmed by
hydrogen-NMR.
[0129] (Step 2):Preparation of
2,3-bis(4-propyloxy-benzylidene)succinaldeh- yde
[0130] 24 mg of 2,3-bis-(4-hydroxy-benzylidene)succinaldehyde(0.08
mmol) prepared in the above step 1 was added to the 100 ml
round-bottomed flask. Thereafter, 30 mg of potassium carbonate(0.18
mmol) and 30 mg of 1-iodopropan(0.18 mmol) were added to the flask.
Thereafter, the mixture was stirred at 85.degree. C. for 12 hours.
After the termination of the reaction is confirmed with thin layer
chromatography, the reaction mixture was filtered to obtain the
filtrate. The residue obtained by concentrating the filtrate was
purified with silica gel column chromatography to prepare 23 mg of
the title compound.
[0131] mp: 152.about.153.degree. C. Rf: 0.5
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.1.01 (t, 6H), 1.73.about.1.84 (m, 4H), 3.90 (t, 4H),
6.80.about.6.83 (dd, 4H), 7.48.about.7.51 (dd, 4H), 7.63 (s, 1H),
9.63 (s, 2H).
EXAMPLE 29.about.34
[0132] In the Example 28, alkylating agent containing R.sub.3 shown
in table 1 was used in place of 1-iodopropan, and the same
procedure as the example 28 was accomplished.
EXAMPLE 29
2,3-bis-(4-isopropyloxy-benzylidene)succinaldehyde
[0133] mp:200.about.202.degree. C. Rf=0.45
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.1.31(d, 12H, CH.sub.3--CH--CH.sub.3), 4.53.about.4.57(m, 2H,
CH.sub.3--CH--CH.sub.3), 6.79(d, 4H), 7.49(d, 4H), 7.62(s, 2H,
CH.dbd.C--), 9.62(s, 2H, CH.dbd.O).
EXAMPLE 30
2,3-bis-(4-valeryoxy-benzylidene)succinaldehyde
[0134] mp: gel Rf=0.25 (Hexane:Ethylacetate=7:3).
.sup.1H-NMR(CDCl.sub.3/T- MS) (ppm): .delta.0.96(t, 6H,
CH.sub.2--CH.sub.3) , 1.37.about.1.49(m, 4H, CH.sub.2--CH.sub.3),
1.64(1.77(m, 4H, CH.sub.2--CH.sub.2), 2.55(t, 4H,
OCH.sub.2--CH.sub.2), 7.04(d, 4H), 7.52(d, 4H), 7.69(s, 2H,
CH.dbd.C--), 9.66(s, 2H, CH.dbd.O).
EXAMPLE 31
2,3-bis-(4-benzyloxy-benzylidene)succinaldehyde
[0135] mp:119.about.120.degree. C. Rf=0.65
(Hexane:Ethylacetate=4:6). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.5.06(s, 4H, O--CH.sub.2-ph), 6.89.about.7.54(m,
18H.sub.arom), 7.64(s, 2H, CH.dbd.C--), 9.64(s, 2H, CH.dbd.O).
EXAMPLE 32
2,3-bis-(4-benzoyloxy-benzylidene)succinaldehyde
[0136] mp:207.about.208.degree. C. Rf=0.36
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.7.21.about.8.18(m, 18H.sub.arom), 7.73(s, 2H, CH.dbd.C--),
9.69(s, 2H, CH.dbd.O).
EXAMPLE 33
2,3-bis-(4-(2-fluorobenzoyloxy)benzylidene)succinaldehyde
[0137] mp:197.about.199.degree. C. Rf=0.31
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.7.17.about.8.09(m, 16H.sub.arom), 7.73(s, 2H, CH.dbd.C--),
9.69(s, 2H, CH.dbd.O).
EXAMPLE 34
2,3-bis-(4-(4-bromobenzoyloxy)benzylidene)succinaldehyde
[0138] mp:223.about.225.degree. C. Rf=0.35
(Hexane:Ethylacetate=6:4). .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.7.19.about.8.03(m, 16H.sub.arom), 7.72 (s, 2H, CH.dbd.C--),
9.69(s, 2H, CH.dbd.O).
EXAMPLE 35
2,3-bis-(2-(N-methylpiperazine) benzylidene)succinaldehyde
[0139] 10
[0140] (Step 1) :2- (N-methylpiperazine)benzaldehyde
[0141] 42.16 ml of 2-fluorobenzaldehyde (400 mmol) was added to the
250 ml round-bottomed flask, 100 ml of dimethylformamide(DMF) was
further added therein. Thereafter, 8.28 g of potassium carbonate
(60 mmol) and 44.4 ml of N-methylpiperazine (400 mmol) were added
therein. The reaction mixture was refluxed at 150.degree. C. for 10
hours.
[0142] After the reaction was terminated, the reaction mixture-was
cooled to the room temperature. Thereafter, 400 ml of water was
added to the reaction mixture. The reaction mixture was extracted
with 200 ml of ethyl acetate for three times. The ethyl acetate
layers were collected, washed with water for three times and dried
with anhydrous magnesium sulfate. The ethyl acetate was removed
under reduced pressure. The resulting residue was purified with
column chromatography, to give
2-(N-methylpiperazine)benzaldehyde.
[0143] (Step 2):Preparation of
2,3-bis-(2-(N-methylpiperazine)benzylidene)- succinaldehyde
[0144] 4.5 g of 2-(N-methylpiperazine)benzaldehyde (30 mmol)
prepared in the above step was added to the 250 ml round-bottomed
flask. Thereafter, 2 ml of 2,5-dimethoxytetrahydrofuran(15 mmol), 2
g of potassium acetate (20 mmol), 1 ml of acetic acid(16 mmol) and
1 ml of water were added to the flask. The reaction mixture was
refluxed at 110.degree. C. for 12 hours. After the reaction was
terminated, the reaction mixture was cooled to the room
temperature. Thereafter, water was added to the reaction mixture.
The reaction mixture was extracted with 100 ml of chloroform for
three times. The chloroform layers were collected, washed with
water for three times and dried with anhydrous magnesium sulfate.
The chloroform was removed under reduced pressure. The resulting
residue was purified with column chromatography, to give the title
compound as a yellow crystal (4.5 g, yield: 65%)
[0145] mp:218.about.220.degree. C. Rf: 0.3 (CHCl.sub.3:MeOH=8:2).
.sup.1H-NMR(CDCl.sub.3/TMS) (ppm): .delta.2.25(s, 6H, OCH.sub.3),
2.40.about.2.85(BRD, 16H, N--CH.sub.2--CH.sub.2--N), 6.94(dt, 2H,
J=1.2, 7.5 Hz), 7.04(dd, 2H, 2H, J=1.2, 7.5 Hz), 7.22(dd, 2H,
J=1.2, 7.8 Hz), 7.32(dt, 2H, J=1.2, 7.5 Hz), 7.92(s, 2H,
CH.dbd.C--), 9.70(s, 2H, CH.dbd.O).
EXAMPLE 36.about.39
[0146] In the Example 1, benzaldehyde substituted with R.sub.1,
R.sub.2 and R.sub.3 shown in table 1 was used in place of
benzaldehyde, and the same procedure as the example 1 was
accomplished.
EXAMPLE 36
2,3-bis-(3-(4-chlorobenzyloxy)-benzylidene)succinaldehyde
[0147] mp:193.about.194.degree. C. Rf: 0.48
(Hexane:Ethylacetate=6:4). .sup.1H-NMR (CDCl.sub.3/TMS) (ppm):
.delta.4.93 (dd, 4H, CH.sub.2-Arom), 6.96.about.7.32(m,
16H.sub.arom), 7.55(s, 2H, CH.dbd.C--), 9.53(s, 2H, CH.dbd.O).
EXAMPLE 37
2,3-bis-(3-(4-methoxybenzyloxy)-benzylidene)succinaldehyde
[0148] mp:117.about.119.degree. C. R.sub.f: 0.38
(Hexane:Ethylacetate=6:4)- . .sup.1H-NMR(CDCl.sub.3/TMS) (ppm):
.delta.3.79(s, 6H, --OCH.sub.3), 4.88(dd, 4H, CH.sub.2-Arom),
6.85.about.7.27(m, 16H.sub.arom), 7.57(s, 2H, CH.dbd.C--), 9.55(s,
2H, CH.dbd.O).
EXAMPLE 38
2,3-bis-(4-(4-chlorobenzyloxy)-benzylidene)succinaldehyde
[0149] mp:208.about.209.degree. C. R.sub.f: 0.44
(Hexane:Ethylacetate=6:4)- . .sup.1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.5.02(s, 4H, CH.sub.2-Arom), 6.86.about.7.52(m,
16H.sub.arom), 7.63(s, 2H, CH.dbd.C--), 9.63(s, 2H, CH.dbd.O).
EXAMPLE 39
2,3-bis-(4-(4-methoxybenzyloxy)-benzylidene)succinaldehyde
[0150] mp:218.about.220.degree. C. R.sub.f: 0.25
(Hexane:Ethylacetate=6:4)- . .sup.1H-NMR(CDCl.sub.3/TMS)(ppm):
.delta.3.81(s, 6H, --OCH.sub.3), 4.97(s, 4H, CH.sub.2-Arom),
6.88.about.7.52(m, 16H.sub.arom), 7.63(s, 2H, CH.dbd.C--), 9.63(s,
2H, CH.dbd.O).
[0151] Cinnamaldehyde derivatives of the present invention prepared
in the examples were shown in table 1.
1TABLE 1 No. R.sub.1 = R.sub.1' (ortho) R.sub.1 = R.sub.1' (meta)
R.sub.1 = R.sub.1' (para) m.p. (.degree. C.) 1 H H H 150-151 2 --F
H H 141-142 3 --Cl H H 177-178 4 --Br H H 194-195 5 --OCH.sub.3 H H
163-164 6 H --Cl H 107-108 7 H --OCH.sub.3 --OH 177-178 8 H
--OCH.sub.3 --OCH.sub.3 149-150 9 H H --Cl 172-173 10 H H --OH
244-246 11 H H --CH.sub.3 177-178 12 H H --OCH.sub.3 204-207 13
--OCH.sub.2CH.sub.2CH.sub.3 H H 109-110 14 --OCH.sub.2CHCH.sub.2 H
H 117-118 15 --OCH(CH.sub.3).sub.2 H H 109-110 16
--OCH.sub.2C.sub.6H.sub.5 H H 120-121 17
--OCH.sub.2C.sub.6H.sub.4-4-Cl H H 186-187 18
--OCH.sub.2C.sub.6H.sub.4-4-Br H H 210-211 19
--OCH.sub.2C.sub.6H.sub.4-4-NO.sub.2 H H 116-117 20 H
--OCH.sub.2CH.sub.2CH.sub.3 H 93-94 21 H --OCH(CH.sub.3).sub.2 H
116-117 22 H --OCH.sub.2C.sub.6H.sub.5 H 126-127 23 H --OCH.sub.3
--OCH.sub.2CH.sub.2CH.sub.3 95-96 24 H --OCH.sub.3
--OCH(CH.sub.3).sub.2 119-120 25 H --OCH.sub.3 --OCOCH.sub.3
149-151 26 H --OCH.sub.3 --OCO(CH.sub.2).sub.3CH.sub.3 gel 27 H
--OCH.sub.3 --OCOC.sub.6H.sub.5 107-110 28 H H
--OCH.sub.2CH.sub.2CH.sub.3 152-153 29 H H --OCH(CH.sub.3).sub.2
200-202 30 H H --OCO(CH.sub.2).sub.3CH.sub.3 gel 31 H H
--OCH.sub.2C.sub.6H.sub.5 119-120 32 H H --OCOC.sub.6H.sub.5
207-208 33 H H --OCOC.sub.6H.sub.4-2-F 197-199 34 H H
--OCOC.sub.6H.sub.4-4-Br 223-225 35 N- H H 218-220 methylpiperazine
36 H --OCH.sub.2C.sub.6H.sub.4-4-Cl H 193-194 37 H
--OCH.sub.2C.sub.6H.sub.4-4-OCH.sub.3 H 117-119 38 H H
--OCH.sub.2C.sub.6H.sub.4-4-Cl 208-209 39 H H
--OCH.sub.2C.sub.6H.sub.4-4-OCH.sub.3 218-220
EXPERIMENTAL EXAMPLE 1
Cell Cycle Analysis
[0152] In order to confirm that the cinnamaldehyde derivatives of
the present invention could regulate cell cycle, the present
inventors performed the following experiment.
[0153] Colon cancer cell of human SW620(ATCC) or HCT116(ATCC) was
transferred to T25 flask(culture 7.5 ml) by RPMI1640(Gibco/BR)
medium or DMEM medium(Gibco/BR) containing 10% FBS (fetal bovine
serum) , respectively. Thereafter the cell was incubated for 12
hours at 37.degree. C. in incubator. After the cell was incubated
for 12 hours, 7.5 .mu.l of DMSO was added to a culture used as
control group, for the final concentration to be 0.1%. In order to
use in experiment, samples of various concentrations were prepared
by dissolving the cinnamaldehyde derivatives of the present
invention in DMSO. 7.5 .mu.l of the prepared samples was added to a
culture used as experimental group. Cell of control or experimental
group was incubated for 48 hours.
[0154] For cell cycle analysis, incubated cell was removed from
medium. Thereafter the incubated cell was separated from the flask
by trypsin. Thus the cell was centrifuged at 300 g for 5 min. The
cell was washed with phosphate buffer solution (PBS) to remove the
residue medium from the cell. 3 ml of ethanol (70%) was added to
the cell. The cell was fixed by being positioned at -20.degree. C.
for 12 hours. The fixed cell was centrifuged at 300 g for 3 min and
washed with the cold PBS for two times to remove residual ethanol.
The cell solution prepared by adding 500 .mu.l of PBS to the cell
was well mixed to produce the homogeneous state. 100 .mu.g/ml RNase
A 50 .mu.l was added to the cell solution, thereafter the resulting
cell solution was positioned at 37.degree. C. for 30 min. The
chromosomal DNA was stained by further adding 1 mg/ml propidium
iodide (soluble in PBS) thereto.
[0155] Distribution of cell cycle of 20,000 stained cells were
measured by using Becton-Dickinson FACS caliber (San Jose, Calif.,
USA). Amounts of cells dividing to G1, S or G2/M of cell cycle was
measured by using Becton-Dickinson Modifit cell cycle analysis
program, resulted in percentage.
[0156] The results were shown in table 2.
[0157] HCT116 GI.sub.50(.mu.M) and SW620 GI.sub.50(.mu.M) mean to
be the concentrations at which growth of HCT116 and SW620 colon
cancer cell was inhibited to the 50%, respectively.
2TABLE 2 G2/M G2/M HCT116 SW620 at 1 .mu.M(%) at 1 .mu.M(%) No.
GI.sub.50(.mu.M) GI.sub.50(.mu.M) SW620 HCT116 1 8.39 1.52 45 42 2
4.02 10.73 58 48 3 0.91 0.91 47 40 4 1.05 2.14 36 38 5 3.72 1.55 50
55 6 3.62 90.58 66 60 7 7.87 3.53 38 30 8 5.49 10.46 75 71 9 3.62
24.16 67 63 10 1.36 1.02 65 61 11 1.03 5.17 44 40 12 9.62 10.24 57
59 13 5.28 2.11 62 65 14 5.28 2.64 76 72 15 5.34 5.34 69 70 16 1.41
0.63 80 43 17 0.84 0.54 76 71 18 2.63 4.64 70 75 19 5.19 7.09 39 32
20 2.74 6.74 76 72 21 4.23 0.61 74 75 22 7.59 3.58 65 68 23 0.62
2.28 56 50 24 0.71 0.91 55 48 25 17.68 66.31 48 43 26 11.18 54.04
47 46 27 2.95 13.87 77 79 28 0.82 0.55 70 72 29 0.82 1.06 47 49 30
20.98 65.05 65 63 31 3.58 7.38 62 64 32 11.94 5.97 52 56 33 7.43
5.57 71 72 34 30.42 16.66 62 60 35 5.23 0.72 79 52 36 7.5 6.3 42 41
37 3.4 5.1 46 35 38 3.8 2.2 53 44 39 3.7 6.8 48 38
[0158] As shown in the table 2, from the analysis of cell cycle of
colon carcinoma cell line SW620 or HCT116, in which the compound of
the present invention was administered, the increase of the number
of a cell was confirmed in G2/M stage. So, the compound of the
present invention is believed to arrest the cell in the stage of
G2/M selectively.
EXPERIMENTAL EXAMPLE 2
[0159] In order to confirm that the cinnamaldehyde derivatives of
the present invention could inhibit the growth of cancer cell, the
present inventors performed the following experiment.
[0160] SW620 or HCT116 as colon cancer cell was inoculated in 100
mm plate to the amount of 2.times.10.sup.6 cells. Then,
cinnamaldehyde derivative of the present invention (compound 16 or
compound 35) was added to the plate in the concentration of 1
.mu.M. Thereafter, the colon cancer cell was incubated for 48
hours. Trypsin was added to the plate, to separate the colon cancer
cell from the plate. And 5 ml of PBS (phosphate buffer solution)
was added to the plate, to prepare the cell mixture. The cell
mixture was centrifuged at 1,500 rpm for 5 min for two times, to
remove trypsin from the mixture. The residue was washed with PBS
for two times, thus the resulting supernatant was discarded. 1 ml
of PBS and 3.about.5 ml of cool ethanol(70%) was added to the
resulting residue. The cell was fixed by being positioned at
-20.degree. C. for 12 hours. The fixed cell was centrifuged at
1,500 rpm for 5 min and washed with the cool PBS for two times to
remove residual ethanol. RNase(10 .mu.g/ml) was added to the fixed
cell, to remove RNA. And DNA of the fixed cell was dyed with
PI(propidium iodide, 50 .mu.g/ml). After the fixed cell was
positioned at 37.degree. C. for 30 min, cell cycles of 20,000 cells
were measured by using Becton-Dickinson FACS caliber (San Jose,
Calif., USA). And amount of cell dividing to G0-G1, S or G2/M of
cell cycle was measured by using Becton-Dickinson Modifit cell
cycle analysis program, resulted in percentage.
[0161] For control group, the same procedure was accomplished
except that DMSO was used in place of the cinnamaldehyde
derivative.
[0162] The results were shown in table 3.
3 TABLE 3 Amount of cell (%) Cell Group Con. G0-G1 S G2-M HCT116
Control 1 .mu.M 34.22 44.63 24.16 Compound 16 1 .mu.M 22.36 34.35
43.29 Compound 35 1 .mu.M 15.52 32.37 52.11 SW620 Control 1 .mu.M
47.45 39.57 12.97 Compound 16 1 .mu.M 6.64 13.15 80.21 Compound 35
1 .mu.M 8.74 12.73 78.53
[0163] As shown in table 3, the compound 16 and the compound 35 of
the present invention held a cancer cell in the stage G2/M. So, the
compounds of the present invention can be effectively used as an
inhibitor of cancer cell growth or a regulator of cell cycle.
[0164] Industrial Applicability
[0165] As explained hereinbefore, the compound of the present
invention shown in the above formula 1 can be used as a cancer cell
growth inhibitor or a cell cycle regulator since it can regulate
the cell cycle by holding a cell in the middle of division in the
stage G2/M and can inhibit the cancer cell growth thereby.
* * * * *